WELCOME TO THE DNA REPAIR INTEREST GROUP, WHERE WE HAVE NEXT MONTH DAVID BOREN WILL BE SPEAKING FROM UNIVERSITY OF KENTUCKY IN JANUARY AND FEBRUARY AND MARCH, WE'RE OPEN FOR SUGGESTIONS. YOU CAN LET WILL OR ME KNOW WHAT'S HAPPENING. AND IN APRIL, NATALIE GASMAN FROM THE UNIVERSITY OF SOUTH ALABAMA WILL BE SPEAKING ABOUT THE RADIATION TO MONITOR SINGLE BREAK STRAND REPAIR. SHOULD BE VERY INTERESTING. WE HAVE AN OPENING IN MAY, WE MAY DO THE YOUNG INVESTIGATOR TALKS THEN, AND IN JUNE, SCOTT WILLIAMS AT NIEHS WILL SPEAK. SO TODAY, IF WE GO BACK TO OUR PURPLE SCREEN, TODAY WE WILL HAVE THE TALK FROM DR.MARK KELLEY, DEVELOPMENT OF THE FIRST CLINICAL DRUG TARGETING DNA REPAIR REDOX SIGNALING. AND WE'LL TALK ABOUT CLINICAL IMPLICATION FOR PANCREATIC AND OTHER CANCERS. LET'S MUTE AT ALL THE OTHER SITES AND TAKE IT AWAY IN BALTIMORE. >> MUTE SO YOU DON'T HEAR ANY ECHO. WE HAVE MARK HERE AND DAVE, BOTH ARE GOING TO SAY A FEW WORDS OF INTRODUCTION. >> YES. GREAT PLEASURE TO GO AHEAD AND INTRODUCE DR.MARK KELLEY WHO IS GOING TO BE SPEAKING TODAY. MARK GOT HIS PH.D. [INAUDIBLE]. HE INITIATED [INAUDIBLE] HE WENT ON TO DO HIS POSTDOC-- THIS IS BACK WHEN SEQUENCING WAS NOT SO TRIVIAL. HE ENDED UP THEN GOING IN THE LATE 80S TO START HIS INDEPENDENT CAREER AT UNIVERSITY OF CHICAGO, HE ACTUALLY MENTORED MANY EXCELLENT GRADUATE STUDENTS. HE ALSO INITIATED WORK THERE-- INITIALLY WORKING IN THE AREA-- >> EXCUSE ME, COULD YOU GET CLOSER TO THE MICROPHONE? YOU KEEP GETTING LOUDER AND SOFTER AND IT'S HARD TO HEAR. >> SORRY ABOUT THAT. HOW ABOUT THIS? >> SPEAK UP, DAVE. >> SO HE WENT TO LOYOLA UNIVERSITY IN THE LATE 80S, [INAUDIBLE] AND ONE OF THE EARLY INVESTIGATORS WHO PIONEERED THE IDEA OF [INAUDIBLE] FUNCTION-- ACTIVITY. HE ALSO INITIATED HIS WORK THERE ON APE1 AND-- NUCLEASES AND STARTED TO PERCEIVE THIS CONCEPT THAT YOU COULD ACTUALLY [INAUDIBLE] BACK WHEN HE WAS AT LOYOLA AND EVENLY-- HE STARTED LOOKING AT-- [INAUDIBLE] SMALL MOLECULES ACTUALLY IN TARGET-SPECIFIC-- OR REDOX ACTIVITIES. SO HE WENT TO INDIANA AGAIN IN THE LATE '90S, WHERE HE CONTINUED HIS WORK IN DNA REPAIR AND REALLY STARTED TO BLOSSOM INTO FOCUSING LARGELY ON [INAUDIBLE]. A GREAT DEAL OF HIS WORK HAS FOCUSED AROUND APE1 AND HE'S DONE A LOT OF THE WORK LOOKING AT THE REDOX FUNCTION [INAUDIBLE] AND I THINK THAT'S WHAT HE'S GOING TO BE TALKING ABOUT, HIS WORK WITH REGARDS TO REF-1. MARK HAS OVER 200 PUBLICATIONS ON PUBMED, SEVERAL AWARDS AND ACCREDITATIONS BUT I'M NOT GOING TO TAKE ANY MORE TIME [INAUDIBLE] >> WELCOME, MARK. >> THANKS. I'M GOOD. THANK YOU, DAVE, THANK YOU, WILL, THANK YOU ALL FOR ATTENDING HERE AND AT THE REMOTE LOCATIONS. IT'S ALWAYS A PLEASURE TO COME UP HERE, IT'S ALWAYS FUN, THESE VISITS, BECAUSE I'M TALKING TO PEOPLE WHO KNOW WHAT I'M TALKING ABOUT AND I DON'T HAVE TO EXPLAIN ALL THE BACKGROUND, AND IT'S ALWAYS WONDERFUL TO VISIT WITH OLD COLLEAGUES AND FRIENDS. >> [INAUDIBLE] >> GOOD POINT. THANKS FOR THE NICE INTRODUCTION. IT'S ALWAYS GREAT TO COME BACK. ONE OF THOSE GREAT GRAD STUDENTS I DID HAVE WAS DAVE WILSON, AS YOU ALL KNOW, SO HE'S BEEN STELLAR AND I'M VERY PROUD. I WISH I COULD TAKE CREDIT FOR IT BUT I CAN'T. ANYWAY, SO WHAT I'D LIKE TO TALK TO YOU ABOUT TODAY IS APE1/REF-1 AS A CLINICAL TARGET. IF I HAVE TIME-- THE FIRST PART OF MY TALK IS GOING TO BE TALKING ABOUT ITS ROLE IN CANCER AND HOW WE'RE TARGETING IT AND BRINGING THE DRUG TO TRIAL, HOPEFULLY IN EARLY 2017, AND THE LAST PART OF MY TALK, DEPENDING ON THE TIME, THE OTHER ASPECTS THAT ARE INVOLVED IN CHEMOTHERAPY INDUCED PERIPHERAL NEUROPATHY AND HOW YOU CAN TARGET APE FROM BOTH KILLING TUMORS AND PROTECTING AGAINST THE CHEMOTHERAPY-INDUCED PERIPHERAL NEUROPATHY. SO I HAVE TO GIVE MY DISCLOSURES. WE'VE ALREADY HAD A MAC TO PC SWITCH. TO I DO HAVE TO DISCLOSE THAT I AM CHIEF SCIENTIFIC OFFICER OF HEXION PHARMACEUTICALS, RECENTLY CHANGED THE NAME, NUMBER OF LICENSING AGREEMENTS. MAKE SURE YOU USE THE APE BODY BECAUSE BUY IT FROM-- THEN WE CAN GET MORE FUNDING FOR MY LAB. THIS IS THE GROUP OF PEOPLE I'M GOING TO BE TALKING ABOUT, MOST OF THE WORK HAS BEEN DONE WITH-- SHE'S BEEN DOING A LOT OF THE-- WORK AND THE REST IS WORKING ON SOME OF THE NEUROPATHY AND PANCREATIC. IN THIS CROWD, HERE IN THIS AUDIENCE, I REALLY DON'T HAVE TO GO INTO THIS TOO MUCH, BUT I NEED TO GIVE A LITTLE BACKGROUND THAT AS YOU KNOW, OR SHOULD KNOW, IT HAS TWO NAMES AND TWO MAJOR FUNCTIONS ALL THOSE THERE'S SOME OTHER FUNCTIONS ATTRIBUTED TO IT INCLUDING THE RNA DEGRADATION AND QUALITY CONTROL FUNCTION, BUT THE TWO THAT WE FOCUS ON A LOT ARE THE REDOX FUNCTION THIS, IS A REDOX SIGNALING FUNCTION WHERE THIS PROTEIN CONVERTS OXIDIZED TRANSCRIPTION FACTORS TO REDUCE STATES SO THEY COMBINE WITH THE DNA AND TURN ON DOWNSTREAM GENES AND THE PROTEINS WE FOCUS ON A LOT AS TARGETS ARE HIP 1-- AND NF KB, ALTHOUGH OTHERS-- THERE'S OBVIOUSLY THE DNA REPAIR FUNCTION WHICH IS INVOLVED IN THE REPAIRING OF AP SITES-- FOLLOWED BY-- IT'S BEEN SHOWN THAT APRIL ALTERED LEVELS MAINLY THE PROTEIN LEVEL IN THE LARGE NUMBER OF CANCER, AND A LOT OF PEOPLE INCLUDING PEOPLE IN THIS ROOM AND AT THE REMOTE SITES HAVE PUBLISHED OF APE'S ROLE, I'M STANDING HERE TODAY BECAUSE OF THE TEAM WORK NOT ONLY BY MY GROUP BUT BECAUSE OF A LOT OF YOU PEOPLE IN THE ROOM AND AGAIN AT THE OTHER SITE. WE BUILD ON THE BASE BEING KNOWLEDGE AND WE WOULDN'T BE WHERE WE ARE TO TAKE THIS DRUG TO TRIAL WITHOUT WORK THAT'S BEEN DONE BY OTHERS, SO I THINK THIS HAS REALLY BEEN-- I CAN'T STRESS THAT ENOUGH TO THE GRAD STUDENTS AND POSTDOCS, THAT REALLY IT'S BEEN A TEAM EFFORT. THE NEAT THING ABOUT TARGETING APE IS THAT WHEN YOU TARGET THE REDOX FUNCTION OF IT, YOU CAN HIT-- KIND OF GET AN AMPLIFIER EFFECT BECAUSE YOU CAN ACTUALLY TARGET NON-DRUGGABLE TARGETS, STAT3, NF KB, HIP ONE ALPHA HAVE BEEN TOUGH TO TARGET DIRECTLY, AND WE CAN DO THAT BY REDOX SIGNALING ACTIVITY. SO BRIEFLY YOU KNOW THE ROLE OF APE IN DNA REPAIR, I'LL TALK ABOUT THAT MORE AT THE END, AND THIS REDOX, JUST TO REITERATE, WE'VE DONE A NUMBER OF STUDIES AND I'M GOING TO BRIEFLY GO THROUGH THE BACKGROUND A LITTLE BIT OF WHAT WE THINK IS THE ACTIVE REGION, IT'S NOT REALLY AN ACTIVE SITE, AN ACTIVE REGION AS A PROTEIN INVOLVING THREE CYSTINES SHOWN HERE ON THE LEFT HAND OF THE SLIDE. APE HAS BEEN SHOWN TO BE ALTERED IN A LOT OF THESE CANCER, WE'VE DONE WORK ON A NUMBER OF THESE, BUT A LOT OF THESE, AGAIN, HAVE BEEN DONE BY THOSE PRESENT IN THE ROOM ALSO AROUND THE WORLD WHO HAVE LOOKED AT THESE DIFFERENT CANCER, AND AGAIN AT THE PROTEIN LEVEL, LOOKING AT THE ROLE OF DYSFUNCTION AT THE PROTEIN LEVEL. A LOT OF TIMES EARLY ON, PEOPLE SAY WHY LOOK AT APE, BECAUSE WE LOOK AT CANCERS AND THE RNA LEVEL DOESN'T CHANGE. AND I THINK THE ANSWER IS, WELL, THAT'S RIGHT, IT DOESN'T CHANGE, IT'S THE FUNCTION, IT'S THE LOCATION, IT'S THE MOVEMENT BETWEEN THE NUCLEUS, THE MITOCHONDRIA, THAT'S WHAT WE REALLY FOCUS ON. SO THESE ARE A LARGE NUMBER OF CANCERS WHERE, AGAIN, IT'S BEEN ALTERED, SO AGAIN IT'S NOT A ONE-TRICK PONY, NOT A CANCER. IT'S VERY IMPORTANT HOW WE USE IT AS A CHEMICAL TOOL. SO IN THE 90S, TOM CONCERN WAS THE FIRST TO SHOW THERE'S A PROTEIN INVOLVED IN REDOX SIGNALING, REDOX FUNCTION, AND HE SHOWED WHEN THEY GOT DONE CLONING AND SEQUENCING, IT WAS REALLY APE-1, SO IT GOES BY APE-1-REF-1. WE WERE INTERESTED IN DNA REPAIR FOR YEARS. WHEN THIS BECAME KNOWN, WE STARTED LOOKING AT THIS ON GENETIC MUTATIONS. BECAUSE YOU CAN MUTE APE AND KEEP THE REPAIR, YOU CAN MUTATE THE REPAIR FUNCTION OF APE AND KEEP THE REDOX. SO IT'S A VERY INTERESTING PROTEIN. AND WE WANTED SOME CHEMICAL MOLECULES TO BLOCK BECAUSE IT WOULD BE NICE, RATHER THAN KNOCKING DOWN APE, KNOCKING DOWN AND TRYING TO ADD BACK, TO HAVE CHEMICAL INHIBITORS, SO OUR LAB AND ACTUALLY DAVID'S LAB AND A NUMBER OF LABS WILL LOOK FOR REPAIR INHIBITORS FOR YEARS AND YEARS, AND WHILE A NUMBER HAVE BEEN PUBLISHED, THEY REALLY HAVEN'T GONE ANYWHERE, THAT IS A HARDER NUT TO CRACK THAN WE THOUGHT IT WOULD BE, WE THOUGHT THAT WOULD BE THE EASY PART. AT THE SAME TIME, THIS MOLECULE IS BEING DEVELOPED IN JAPAN CALLED E33-30, AND IT WAS PART OF A BIG PUSH THEY HAD LOOKING AT NF KAPPA B INHIBITORS, INFLAMMATION IN HEPATITIS. SO AFTER A LARGE BIG PHARMA DEVELOPMENT, THEY CAME UP WITH THIS MOLECULE THAT THEY FOUND SPECIFICALLY BLOCKED NFKB ACTIVITY, AND WHEN THEY DID MORE OF THE BIOCHEMISTRY, IT TURNS OUT IT DOESN'T TOUCH NFKB AT ALL, IT BINDS TO APE. SO THEY THEN WENT ON AND DEVELOPED IT THROUGH A VIGOROUS CLINICAL TRIAL PROGRAM, 10 CLINICAL TRIALS, 422 PATIENTS, ALL IN JAPAN, NEVER PUBLISHED, NEVER REGISTERED, AND AT THE END OF THE TRIAL, THEY ABOUT A 30% EFFICACY THEY SAW WITH THE DRUG WITH CHRONIC HEPATITIS C AND B. PATIENTS WERE TAKING IT UP TO ABOUT 240MILLIGRAMS PER DAY, SOME PATIENTS WENT UP TO 600. THEY WERE ON IT FOR THREE MONTHS WITH VERY LITTLE SIDE EFFECTS. MAYBE A SLIGHT DIARRHEA. VERY SAFE DRUG, VERY TARGETED DRUG, AND ABOUT THE TIME THEY GOT THE DATA, AT THE BOTTOM OF THE SCREEN, THEY END LICENSED TWO DRUGS. REVOVIR AND HUMIRA. IF YOU WATCH ANY TV AT ALL, IT'S ON EVERY OTHER AD. SO WHAT ARE YOU GOING TO DO, TAKE THE OTHERS AND RUN WITH IT, THAT'S WHAT THEY DID. SO THE PATENT IS EXPIRED, AND BECAUSE OF OUR INTEREST IN LOOKING AT SMALL MOLECULE INHIBITORS OF H REPAIR AND REDUCTION, WE GOT INVOLVED AS THE DRUG-- WE ACTUALLY HAD THE DRUG MADE IN SHOW WALTER'S LAB UP IN UNIVERSITY OF MICHIGAN AND SOME OTHER PLACES NOW, SO IT'S BEEN DEVELOPED AND THAT'S THE DRUG WE'VE BEEN WORKING ON. IT HAS THE NEW NAME BECAUSE IT'S BEEN LICENSED, BUT IF YOU LOOK IN THE LITERATURE, YOU CAN LOOK AT E3330OR APX3330. WE HAVE PUBLISHED AND SOME OTHERS HAVE PUBLISHED IT HAS A DIRECT EFFECT ON APE BY A NUMBER OF BIOCHEMICAL METHODS SHOWN IN THE TOP BULLET. IT DID NOT HAVE AB ACCEPTABLE TOXICITY, BOTH IN ANIMAL AND HUMAN STUDIES, IT WAS A VERY RIGOROUS CLINICAL PACKAGE AND THE PRE-CLINICAL DATA SUPPORTS ITS USE NOT ONLY AS AN INDEPENDENT AGENT, WHICH I'LL SHOW YOU, BUT WE REALLY THINK THE FUTURE IS THIS COMBINATION WHERE YOU CAN NOW LOOK AT WHO APE TALKS TO IN DIFFERENT CELLS, NOT ALL CELLS ARE ALIKE, NOT ALL CANCERS ARE ALIKE, NOT EVEN SUBGROUPS OF CANCERS WILL BE THE SAME, AND YOU CAN PREDICT WHAT WOULD BE GOOD POTENTIAL COMBINATION, AND THAT'S WHAT I'M GOING TO SPEND A FAIR AMOUNT OF MY TIME ON FOR THE DAY. SO APE HAS THIS REDOX ACTIVE REGION, YOU BLOCK IT WITH THE DRUG, TRANSCRIPTION FACTORS FROM OXIDIZED REDUCED STATE, THEY CAN NO LONGER BIND WITH THE DNA, AND YOU TURN OFF GENES THAT ARE INVOLVED IN GROWTH AND ANGIOGENESIS AND OTHER MIGRATION. THE CYSTINES ARE MAINLY 65, 93 AND 99. THIS IS A NUMBER OF PUBLICATIONS, ACTUALLY MY NAME IS ON THE PAPERS BUT A LOT OF THE CREDIT DEPOSE TO MILLI WHO DID A LOT OF THIS WORK, ACTUALLY DID SOME ZEBRAFISH I'LL SHOW YOU. THESE ARE THE THREE THAT ARE REALLY IMPORTANT, THEY FORM SULFIDE BONDS WITH ONE ANOTHER. THE INTERESTING THING IS, WE WERE STUDYING-- A GRANT LOOKING AT ZEBRAFISH APE BECAUSE ZEBRAFISH HAS SIX OF THE SEVEN CYSTINES, APE HAS 7, 3 ARE INVOLVED IN REDOX, MAINLY CYSTINE 65, TURNS OUT ZEBRAFISH HAVE SIX OF THE SEVEN AND THE ONE MISSING IS-- 58 IN ZEBRAFISH THAT IS NOT A CYSTINE, AND ZEBRAFISH APE HAVE FULL DNA REPAIR ACTIVITY, NO REDOX ACTIVITY, AND WE ACTUALLY PUBLISHED YEARS AGO THAT IT CONVERTS-- TO CYSTINE, IT GAINS REDOX FUNCTION. SO IT'S A GAIN OF FUNCTION EVOLUTIONARY CHANGE THAT'S OCCURRED IN THE MOLECULE AND IT'S NOT ENOUGH TO BE A VERTEBRATE, YOU HAVE TO BE A FURRY MAMMAL. SO VERY INTERESTING HOW THIS EVOLVED. WE HAVEN'T REALLY DONE MUCH MORE ON THE EVOLUTION. THERE'S MORE PUBLICATIONS ON THIS THAN THAT, BUT IT'S VERY INTERESTING WHY THIS HAS OCCURRED. WE DID A NUMBER OF STUDIES, WE'VE SHOWN THE DRUG WE HAVE, IF YOU TAKE ALL OF THE CYSTINES OUT OF APE AND ADD BACK TWO OR THREE IN DIFFERENT COMBINATIONS, IT'S ALWAYS C65, YOU HAVE TO HAVE THE 65TO BE ACTIVE. WE SEE THAT THE DRUG BLOCKS THE ACTIVITY REGARDLESS OF WITH THE MISSING FOUR CYSTINES, SO IT'S REALLY THESE THAT ARE IMPORTANT IN ITS REDOX FUNCTION INTERACTING WITH ONE ANOTHER. THE WAY IT WORKS, IF YOU LOOK AT THE TOP PANEL A AND GO LEFT TO RIGHT, YOU'LL SEE THAT IF YOU TAKE APRIL AND YOU-- IN THE END, AFTER SIX HOURS, YOU GET JUST THE SURFACE CYSTINE BUT GET LABELED WITH THE DRUG. WHAT THE MOLECULE IS REALLY DOING IS IT'S INTERACTING WITH CYSTINE 65AND OVER TIME, THE MOLECULE SLOWLY UNFOLDS. THIS IS PART OF HOW IT FUNCTIONS. SO IT'S A VERY DIFFERENT INTERACTION, VERY DIFFERENT INHIBITORY EFFECT THAT YOU GET USING THIS MOLECULE THAN YOU LOOK AT, LIKE, WITH THE KINASE OR A PHOSPHATASE. SO YOU'RE BLOCKING PROTEIN-PROTEIN INTERACTION AND YOU'RE CAUSING THIS UNFOLDING, AND THIS IS SHOWN HERE IN THE MIDDLE PANEL OF THIS SLIDE ON THE RIGHT, WHERE THE END KIND OF UNFOLDS AND THEN OVER TIME IT FOLDS. I JUST PUT THIS UP HERE THAT IF YOU'RE INTERESTED IN READING MORE ABOUT THE MECHANISM, IT WAS IN THIS PUBLICATION MAINLY BY MILLI. SO WHAT IT DOES IS IT'S DESTABILIZING THE STRUCTURE, BUT THEN OVER TIME, IT CAN REFOLD BACK ONCE 3330 IS GONE. SO WHEN YOU START TO THINK ABOUT THIS FROM A PK-PD DYNAMICS AS A DRUG, IT'S GOING TO BE VERY DIFFERENT BECAUSE YOU'RE GOING TO GET THE SLOW UNFOLDING OF THE PROTEINS, THEN YOU'RE GOING TO GET A COMING BACK TOGETHER. SO THAT AFFECTS KIND OF THE DOSING YOU NEED AND THE TREATMENT YOU NEED OF THE DRUG. WE'VE DONE-- AND AGAIN, I'M SORRY I'M JUST GIVING YOU LITTLE SNIPPETS AS A BACKGROUND, BUT WE'VE DONE A LOT OF STUDIES WHERE WE'VE KNOCKED DOWN APE-- THIS IS AN EXAMPLE WHERE WE ADD BACK APE AND HAVE THE DRUG SHOWING THAT JUST BY ADDING THE PROTEIN BACK, YOU HAVE TO USE MORE DRUGS, WHICH IS A CLASSIC TYPE EXAMPLE IN DRUG DEVELOPMENT TO SHOW THAT YOU ARE TYPE EXAMPLE IN DRUG DEVELOPMENT TO SHOW THAT YOU ARE ON TARGET. WE HAVEN'T LOOKED AT EVERY SINGLE ONE BUT WE HAVE LOOKED AT-- THE DRUGS TO NOT HIT THAT. IT'S VERY SPECIFIC FOR APE, IT'S VERY SPECIFIC FOR HITTING THE CYSTINE 65. WE'VE DONE A NUMBER OF STUDIES THAT SOLIDIFY THIS CONCEPT, SO BY PUTTING IN REPORTERS IN VARIOUS CELLS, AND A LOT OF THIS HAS BEEN DONE IN PANCREATIC CANCER, YOU PUT IN THE NF KB OR STAT3, YOU CAN THEN SHOW YOU GET A DOSE RESPONSE OF THE BLINDING OF THESE FACTORS TO THE REPORTER IN THE CELLS. NOW IF YOU'LL NOTICE WE'RE IN THE MICROMOLAR RANGE. A LOT OF YOU, IF YOU'RE MEDICINAL CHEMISTS YOU'RE FREAKING OUT. AGAIN, A COUPLE THINGS. FIRST OF ALL WHEN THEY DID THEIR TRIALS, THEY SAW RANGES IN THE PATIENTS FROM 25 UP TO ABOUT 150 MICROMOLAR LEVEL. THE DRUG WAS VERY SAFE EVEN AT THOSE LEVELS. SO THE DOSING, WHEN WE USE OUR ANIMAL AND ALSO IN OUR CELLS, WE'RE WAY BELOW WHAT'S ACHIEVABLE IN PATIENTS AND WHAT'S EXPECTED. SO WHILE WE'VE BEEN SPENDING A LOT OF TIME LOOKING AT PANCREATIC CANCER, IT'S A TERRIBLE DISEASE, THE FIVE-YEAR SURVIVAL RATE JUST WENT UP TO 7% FROM 5%. THAT'S OVER A 30-YEAR PERIOD SO WE REALLY HAVE MADE VERY LITTLE PROGRESS. IT'S A TOUGH ONE BECAUSE IT'S VERY HYPOXIC, VERY INFLAMMATORY, AND THERE'S A LOT OF MICROENVIRONMENT INVOLVED IN THIS. SO WE SPENT A LOT OF TIME LOOKING AT THE ROLE OF APE REF-1 IN THE TUMOR AND IN THE MICROENVIRONMENT AND OF COURSE BECAUSE OF THE TRANSCRIPTION FACTORS THAT APE HITS AND REGULATES, THE INFLAMMATORY-- MAKES IT A GOOD CANDIDATE. THIS IS MY ONE SLIDE THAT SHOWS THAT HIGH LEVELS OF APE-- THIS IS IN RNA. KEEP IN MIND IN PANCREATIC PATIENTS, USUALLY ABOUT 15 TO MAYBE 20% HAVE THE WHIPPLE SURGERY DONE, SO YOU'RE ONLY LOOKING AT WHAT WAS BANKED-- AND THIS CHANGES ALL THE TIME-- WHAT WAS BANKED OF THE 20% OF PATIENTS WHO CAN HAVE SURGERY. ABOUT 80% OF THE PATIENTS OR MORE DO NOT HAVE-- CANNOT HAVE SURGERY SO YOU'RE ONLY LOOK AT A SUBSET RIGHT AWAY. IN THIS CASE IT SHOWS AT VERY HIGH LEVELS OF APE, WE HAD A LOWER SURVIVAL TIME, ALTHOUGH THE SURVIVAL TIMES AREN'T GREAT EVEN IF YOU HAVE THE LOW LEVEL. WE HAVE SHOWN THAT APE IS HIGHLY EXPRESSED IN THE PANCREATIC TUMOR SAMPLES AND IN THE FIBROBLASTS IN THE MICROENVIRONMENT AROUND IT, AND THAT'S IMPORTANT AS I GO THROUGH MY TALK BECAUSE WE HAVE SWITCHED PRETTY MUCH FROM NOT DOING TUMOR CELLS ON A PLASTIC-- IF WE DO TUMOR CELLS ON PLASTIC, IT'S ALWAYS WITH CANCER-ASSOCIATED FIBROBLASTS FROM THE PATIENT, SO IT'S ALWAYS COLD CULTURE AND WE'RE REALLY MOVING ALMOST COMPLETELY INTO THE 3D-- MODEL BECAUSE YOU CAN ADD-- AND THIS IS MELISSA'S WORK, I'LL SHOW YOU WHERE YOU CAN HAVE TUMOR, CANCER-ASSOCIATED FIBROBLASTS, WE'VE BEEN ABLE TO ADD IN MACROPHAGE, SHE'S ACTUALLY WORKING TO ADD IN EPITHELIAL CELLS TO BRING IN BLOOD VESSELS AND TO REALLY BUILD A 3D STROMA TUMOR WHICH IS GOING TO BE MORE REALISTIC OF THE CANCER RATHER THAN A TUMOR ON PLASTIC. APE EXPRESSED IN POT-- THE LOWER LEFT SHOWS A COUPLE PATIENT DRIVE LINES AND THEN UNDER STROMA, SOME CANCER-ASSOCIATED FIBROBLAST-- SO THIS IS OUR APPROACH. PANC IS A GOOD MODEL BECAUSE THESE TRANSCRIPTION FACTORS ARE INVOLVED AND ACTUALLY STEP 3IS TURNING OUT TO BE VERY IMPORTANT TOO. IT'S INVOLVED BOTH IN THE TUMOR, IT'S INVOLVED IN THE MICROENVIRONMENT, IT'S INVOLVED IN METASTASIS, AND WE FIND APE1 REF-1 LEVELS EXPRESSED AT FAIRLY HIGH LEVELS IN ALL OF THOSE. WE'RE USING ONLY PATIENT-DERIVED LINES, WE ARE USING NO MORE ESTABLISHED CELL LINES. WE'VE PRETTY MUCH MOVED AWAY FROM THOSE. AND THESE ARE PATIENT DRIVE LINE WHERE YOU CAN LABEL THE TUMOR WITH RED AND THE CAFS WITH GREEN, YOU CAN LOOK AT THE HYPOXIC INTERNAL REGION, AND THIS IS KIND OF THE PICTURE YOU GET. THE NEAT THING THIS IS YOU CAN THEN MEASURE, WHEN YOU'RE ADDING DRUGS AND DOING DIFFERENT MODIFICATIONS OF THE RED OR THE GREEN BEING KILLED, BECAUSE THERE'S A DEBATE IN PANCREATIC CANCER THAT YOU DON'T WANT TO KILL TOO MUCH OF THE FIBROBLASTS BECAUSE IT ACTUALLY MAKES THE TUMOR WORSE. HAVING SAID THAT, PAPERS HAVE COME OUT RECENTLY SAYING WE DO WANT TO KILL THE FIBROBLASTS AND THE TUMOR, SO THE JURY IS STILL OUT, SO THAT'S WHY WE'RE LOOKING AT BOTH TYPES. I JUST WANT TO SHOW YOU THIS BECAUSE IN ALL OUR XENOGRAPH MODELS, THERE'S A GENETIC MODEL, COUPLE MODELS IN PANC THAT WE USE, AND THEN IF WE DO ORTHOTOPIC MODELS, THE DIFFERENCE BETWEEN ANTITUMORAL LAW, I THINK THIS SLIDE SHOULD GIVE A LOT OF US PAUSE. IF YOU LOOK AT THE BLUE LINE, THAT'S GROWING THIS PATIENT DERIVED TUMOR CELL BY ITSELF. THE OTHER LINES WE ADD CANCER ASSOCIATED FIBROBLASTS TO THE TUMOR CELLS AND THEN IMPLANT THEM, AND THE GROWTH RATE IS DRAMATICALLY MORE VIGOROUS. SO FRANKLY, IF WE'RE REALLY GOING TO GET AFTER CANCERS LIKE THIS, THIS IS WHAT YOU HAVE TO BE DOING. YOU HAVE TO BE DOING A MODEL THAT IS ROBUST AND IT'S TOUGH TO KILL, PROBABLY NOT THE ONES WHERE YOU'RE GOING TO GET THE BETTER RESULTS IN. SO WE ARE SHIFTING EVERYTHING TO THESE SORT OF ANIMAL MODELS WITH THE TUMOR AND FIBROBLASTS. HERE IS AN EXAMPLE OF WHERE WE TREATED A DRUG IN THE 3D SPHEREOID. THE TOP ROW IS A PRIMARY PANCREATIC LINE, THE PAOC3IS A METASTASIS OF THE PATIENT AND YOU CAN SEE THAT IN A DOSE-RESPONSE, WE GET THE KILLING OF THE TUMOR CELLS MARKED BY THE RED GOING AWAY, AND THAT'S TUMOR ALONE, THEN WE DO IT TUMOR PLUS CAF, AND IT SHOWS IT TAKES EVEN MORE DRUG TO GET THE SAME KILLING IF YOU LOOK AT THIS SLIDE, SAME THING, BUT NOW YOU CAN SEE WHAT YOU'RE GETTING ON THE BOTTOM IS THE CAFS REALLY AREN'T BEING KILLED AS MUCH AS THE TUMOR. SO FOR WHATEVER REASON, BLOCKING THE REDUCTION ACTIVITY OF BLOCK REF-1 IS BLOCKING THE TUMOR GROWTH BUT NOT HAVING A BIG IMPACT ON CAF, SO WE'RE LOOKING AT THIS CROSSTALK BETWEEN THE CAFS AND THE TUMOR, AND HOW TO BLOCK IT IN BOTH DIRECTIONS. DO YOU KNOW ANYTHING ABOUT PANCREATIC CANCER, JIM IS ACTUALLY THE CHAIRMAN OF THE BOARD OF MY COMPANY HOME REPAIR TO DEVELOP-- SOME OTHER DRUGS AT LILI, AND WE WERE WONDERING IF-- WOULD SHOW AN ENHANCED EFFECT AND KILLING. WE DIDN'T JUST DO THIS BLINDLY, OF COURSE THE STANDARD OF CARE IS BECOMING REPLACED BY THE FOFIRINOX PROTOCOL, BUT-- THE REASON WE WANTED TO LOOK AT THIS IS BECAUSE MELISSA PUBLISHED A NICE PAPER IN BIOCHEMISTRY, JBC, I'M SORRY, SHOWING A RELATIONSHIP BETWEEN THE ENZYMES THAT DEGRADE GEMCITABINE, SO WE WANTED TO SEE IF THIS WOULD EXTEND TO OUR 3D TUMOR MODELS. THE BOTTOM LINE IS THAT YOU DO GET A COMBINATION EFFECT WHEN YOU ADD OUR DRUG PLUS GEMCITABINE AS WE PREDICTED FROM THE BIOCHEMISTRY. TUMOR PLUS CAF AND THE RIGHT SIDE ARE CAFS ONLY, WE ALWAYS DO RED AND GREEN, THAT WASN'T DONE FOR CHRISTMAS. IT'S ALWAYS RED AND GREEN. I HAVE TO GET THESE TALKS IN BEFORE THE HOLIDAYS THOUGH. SO WE ALWAYS DO NOW THESE 3D MODELING, AND THEN WE WENT ON AND DID JUST A QUICK STUDY TO SEE IF WE GET THE SAME EFFECT IN VIVO, AND THIS HAS BEEN DONE A COUPLE YEARS AGO SO PREPATIENT LINE, BUT YOU CAN SEE WHEN YOU HAVE THE COMBINATION EFFECT, IF YOU LOOK IN THE LOWER LEFT, YOU CAN SEE WE ONLY TREATED FOR THREE WEEKS AND THEN STOPPED. SO ALL THIS IS WITHOUT TREATMENT, WHICH YOU NORMALLY GO ON, YOU CAN SEE WE HAVE ACTUALLY AT LEAST AN ADDITIVE EFFECT IF NOT OUT LATER ON AS A MORE SYNERGISTIC EFFECT WITH THIS TREATMENT. SO WE WERE GOING BACK AND DOING THESE STUDIES NOW A LITTLE BIT MORE, BUT QUITE FRANKLY WE'RE MORE INTERESTED IN PROBABLY LOOKING AT OUR DRUG AND VARIOUS TARGETED PATHWAY INTERACTIONS RATHER THAN WITH GEMCITABINE. WE'RE NOT THE ONLY ONES THAT LOOKED AT THE DRUG 330IN THE TARGET. THE TOP LEFT IS-- THIS IS YEARS OLD NOW, THIS IS ESTABLISHED CELL LINE, XENOGRAPH WITH THE DRUG, STANDARD 25MGS PER KILOGRAM, YOU IMPLANT THE TUMOR CELLS IN THE PANCREAS, WE LOOKED FOR REDUCTION IN METASTASIS IN THIS MODEL. THE UPPER IS DONE BY A GROUP PUBLISHED LAST YEAR WHERE THEY LOOKED AT 3330 IN COMBINATION WITH 5FU AND THEY SAW SUPPRESSION OF THE COLON CANCER STEM CELLS AND ACTUALLY THEY SAW A VERY DRAMATIC EFFECT AND WE'RE IN THE PROCESS OF REPEATING THIS. THE NEXT SLIDE I'M GOING TO SHOW YOU IS OTHER WORK, I'M NOT GOING TO GO INTO THE DETAILS, BUT OTHER PEOPLE HAVE ALSO USED THE DRUG IN THIS ENDOTHELIAL CELL TUMOR GROWTH MODEL, AND THEY SHOW IN THE LOWER RIGHT THAT ADDING THE DRUG, BLOCKING APE'S REDOX FUNCTION BLOCKED THE TUMOR GROWTH. WHAT'S INTERESTING IS THAT CRT DRUG ON THERE, IF YOU CAN SEE IT, IS NOT A VERY GOOD APE 1REPAIR INHIBITOR, AND ACTUALLY THEY KIND OF PROVE THAT OINT OR IF IT IS A REPAIR INHIBITOR, IT HAD NO EFFECT. SO I'M CAUTIOUS WITH THE SLIDE SO I CAN TELL YOU THAT THE 33 BLOCK-- YOU SEE THAT EFFECT AND I DON'T KNOW IF THAT'S REALLY A GOOD REPAIR INHIBITOR IN VIVO WHERE YOU GET A DUAL EFFECT. IF IT IS A GOOD REPAIR INHIBITOR, THEN IT ADDED NOTHING TO THE TREATMENT, SO IT'S THE REDOX THAT'S GOOD ALONE. SO WHERE ARE WE GOING WITH ALL THIS BACKGROUND? SO WE WERE STARTING A PHASE 1CLINICAL TRIAL IN EARLY 2017 TO IDENTIFY OUR PHASE 1B, WHERE YOU'RE GOING TO FOCUS ON ALL SOLID TREATMENTS, ALL-COMERS IN THE FIRST, WE HAVE 20 TO 40 PATIENTS, BUT WE ANTICIPATE AN ACCELERATED DESIGN, BECAUSE THE DRUG HAS BEEN SHOWN TO BE SAFE, HEPATITIS PATIENTS, WE THINK IT WILL BE A WAY TO ACCELERATE RAPIDLY, PROBABLY ONLY NEED 10 TO 12 PATIENTS, WE PROBABLY WON'T HIT AN-- THE DRUG IS NOT A TOXIC DRUG, AND WE WILL USE BIOMARKERS, PD BIOMARKERS BASED ON HITTING VARIOUS TARGETS THAT MAY BE OUR STUDY END POINT. THE INDS ACCEPTED OBVIOUSLY IN THE IRB, THIS IS ALL JUST A SAFETY TRIAL. I JUST WANT TO PAUSE FOR A MINUTE AND SAY THAT I'M VERY FORTUNATE, AGAIN, LIKE I SAID, BUILD ON THE BASIC SCIENCE OF ALL OF YOU, AND ALSO THAT I'VE HAD A VERY GOOD RELATIONSHIP WITH-- WE'VE HAD GOOD STRONG ACADEMIC SUPPORT THROUGH OUR CANCER CENTER AND WE STARTED A COMPANY TO REALLY TAKE US THROUGH THIS NEXT PHASE, WHICH IS-- AS ANY OF YOU HAVE DONE IT, IS NOT FOR THE FAINT HEARTED TO MOVE A DRUG INTO THEIR TRIAL, GET IT MANUFACTURED, GET IT MADE, GET THE TRIAL, SO THAT'S WITH A VERY GOOD GROUP OF PEOPLE, SO WE HOPE TO START THAT SOON. I ALSO WANT TO POINT OUT AGAIN, WE'RE NOT A ONE TRICK PONY. SO 3330IS COVERED BY A SAFE ORAL DRUG BUT WE'VE DEVELOPED A LARGE NUMBER OF OTHER DRUGS. ONE OF THEM IS APX2009 THAT WE'VE RECENTLY PUBLISHED WITHIN THE LAST FEW MONTHS. WE'VE DONE A VIGOROUS SAR ANALYSIS TO DEVELOP NEW COMPOUNDS, SO THESE ARE PROBABLY TENFOLD MORE POTENT AS WE'RE MOVING FORWARD, SO WE'RE IN THE PHASE NOW OF STARTING TO WORK ON THE FORMULATION AND APD OF THESE MOLECULES IN A PRE-CLINICAL, SO THEY WILL NOT BE FAR BEHIND. I JUST PUT THIS UP HERE TO SHOW YOU THAT EVEN THOUGH I FOCUSED A LOT ON PANCREATIC CANCER ON THE TOP, WE'VE LOOKED AT OTHER CANCERS INCLUDING ALL AND WHAT'S NOT ON HERE IS A PREVENTIVE AGENT FOR AML, AND WE'VE LOOKED AT AND I'M GOING TO TRY TO TALK ABOUT THE CHEMOTHERAPY PERIPHERAL INDUCED PERIPHERAL NEUROPATHY. WE'VE ALSO SHOWN THAT BLOCKING APRIL'S REDOX FUNCTION WILL PARTNER NICELY WITH ANTI-- FOR POTENTIAL USE IN AGE-RELATED MACULAR DEGENERATION, THE WET FORM, AND THEN THE COMBINATIONS WHICH I'M GOING TO TALK MORE ABOUT. SO WE'RE STILL FOLLOWING THE SCIENCE. I'M NOT A CLINICAL PERSON, IT'S REWARDING TO MOVE SOMETHING FROM THE BENCH TO THE CLINIC BUT WE'RE STILL FOLLOWING THE SCIENCE IN THE DIFFERENT CANCERS AND WE HAVE COLLABORATION IN ALL THESE DIFFERENT CANCERS INCLUDING SOME OTHER DISEASES THAT I CAN TALK ABOUT IF WE HAVE TIME. ONE OF THE OTHER NEAT THINGS WE'VE DONE RECENTLY, AND I CAN SAY THAT BECAUSE MELISSA REALLY DID IT, IS WE'VE BEEN ABLE TO TAKE PATIENT DERIVED PANCREATIC TUMOR CELLS AND USE THE FLUIDS, DO SINGLE CELL SORTING SO YOU GET A SINGLE CELL IN A WELL. AND WE DID APE KNOCK DOWN, SCRAMBLED OR KNOCK DOWN MULTIPLE TIMES, THEN WE DID RNA SEQ ANALYSIS. THE BEAUTY OF THIS, THE DATA IS REALLY CLEAN. ALSO IT KEEPS THE BIOINFORMATICS PEOPLE HAPPY. IT'S CLEAN BECAUSE YOU CAN LOOK AT THE KNOCK DOWN AND LOOK AT THE TRANSCRIPTOME YOU GET FROM THAT. SO IF APE IS DOWN 50%, YOU CAN LOOK AT HOW MUCH THE GENES CHANGE, WHICH GENES, IF IT'S DOWN 80%, YOU CAN DO THIS, AND WHAT WE FOUND BY DOING THIS IS THAT A NUMBER OF PATHWAYS CAME UP. ANOTHER NEAT THING IS BECAUSE YOU'RE DOING THE WHOLE GENOME, TRANSCRIPTOME, YOU KNOW, YOU START TO LOOK AT-- YOU'RE NOT JUST LOOKING AT 1STAT3 GENE, WE'RE LOOKING AT EVERYTHING IN THE PATHWAY. SO WHEN GENES COME UP THAT A LOT OF THEM ARE AFFECTED IN THE SAME PATHWAY, YOU START TO BE SUSPICIOUS THAT YOU SHOULD BE LOOKING IN THOSE PATHWAYS, AND LUCKILY OF THEM THAT CAME UP WERE STAT3 AND HIF1, WHICH WE KNEW WERE IMPORTANT, SO WE FELT VALIDATED BY THAT, THEN SOME NEW ONES. WE'RE PREPARING A MANUSCRIPT ON THIS BUT I WANT TO SHOW YOU ONE LITTLE PIECE OF DATA IN THE NEXT SLIDE. SO AFTER YOU DO THE SINGLE CELL RNA KNOCK DOWN, ON THE TOP, PANEL A, THERE'S THREE GENES, THERE WERE CHANGE-- HUGELY CHANGED. YOU CAN SEE BY THE P VALUE HOW MUCH IS LEFT, SO THEY WERE VERY MUCH CHANGED. THE BEAUTY OF THIS IS YOU CAN NOW START TO BUILD YOUR OWN KIND OF ARRAY OR CHIP OR WHATEVER YOU WANT TO CALL IT WHERE YOU CAN LOOK AT-- YOU CAN LOOK AT A LOT OF THEM THAT CHANGE, THEY GO UP AND DOWN. YOU CAN BUILD THIS LITTLE PORTFOLIO OF GENES THAT YOU CAN LOOK AT AND VALIDATE. SO WE WENT BACK AND DID THE SAME KNOCKDOWN IN THE LEFT-HAND SIDE, OF A POPULATION OF THE SAME CELLS USING THE SAME SI RNA AND VALIDATED THAT THE GENES CHANGE IN A POPULATION OF CELLS AS IN SINGLE CELL. AND OF COURSE THE CHANGE WON'T BE AS DRAMATIC AS WHEN YOU DO KNOCK DOWN, BUT THIS IS A GOOD WAY TO VALIDATE. WE ALSO USE IT AS A DRUG VALIDATION, BECAUSE WHEN WE TREAT IT WITH OUR DRUG-- SO THESE GENES THAT I CAN'T TELL YOU ABOUT YET, SORRY, BUT APPEAR TO BE REGULATED THROUGH THE REDOX FUNCTION-- SO WE'RE USING THIS BOTH FOR DISCOVERY IMMUNE PATHWAYS FOR DRUG DEVELOPMENT, SO WHEN WE DEVELOP NEW REDOX INHIBITORS, WE CAN TEST AND SEE IF YOU GET THE SAME PATTERN. IF IT CHANGES, IT MIGHT MEAN YOU'RE OFF TARGET. OR MAYBE SOMETHING ELSE IS GOING ON. BUT IT'S EXCITING TO US BECAUSE IT GIVES US SOMETHING ELSE WE'RE DOING, WE'RE NOW ABLE TO TAKE ALL THE DATA WE DID FROM THAT SINGLE CELL KNOCK DOWN, WE WENT TO OUR BIOINFORMATICS PEOPLE AND STATISTICS PEOPLE AND WE NOW CAN LOOK FOR ALL THE PATHWAYS WE HIT AND ALL THE FDA-APPROVED DRUGS THAT ARE OUT THERE AND OTHER DRUGS THAT ARE BEING DEVELOPED, AND WE'VE BUILT A MATRIX NOW OF POTENTIAL COMBINATION. SO YOU CAN ENVISION THAT IF PATHWAY N IS AFFECTED AND THERE'S A DRUG FOR N AND 3330KNOCKDOWN AFFECTED IT AND 3330KNOCKS IT DOWN AND YOU PUT AN INHIBITOR OF PATHWAY IN WITH OUR DRUG, YOU WOULD GET A COMBINATION OF THAT. SO IT'S GOING TO BE ANOTHER WAY OF SCREENING FOR VIABLE COMBINATIONS AND BECAUSE THE DRUGS ARE FDA-APPROVED, ONCE OUR DRUG HOPEFULLY GOES THROUGH PHASE 1, WE HAVE NOW A WHOLE BUNCH OF NEW INDICATIONS IN COMBINATION THAT WOULD BE POTENTIALLY ABLE TO DO VERY QUICKLY. SO THIS IS A VERY RAPID FROM DISCOVERY TO CLINIC, WE THINK, APPLICATION OF THIS TECHNIQUE. THE OTHER THING YOU CAN DO IS, YOU CAN NOW START LOOKING AT OTHER CANCERS, DOES THIS SAME PATTERN OCCUR WHEN YOU DO THIS, YOU BUILD A PROFILE AND ARE YOU DEVELOPING A PRECISION MEDICINE APPROACH. IF YOU HAVE A CERTAIN PATTERN WITH APE THAT WE SEE WITH THE KNOCK DOWN OR WITH OUR DRUG, IS THAT THEN THOSE PATIENTS WOULD BE THE ONES YOU'D WANT TO TREAT IN THE VARIOUS COMBINATIONS. I HAVE TO BE DONE BY-- SO THE OTHER THING I WANTED TO POINT OUT IS A LITTLE MORE BASIC SCIENCE, THIS IS JUST RECENTLY PUBLISHED IN PANCREATIC CANCER, WE REALLY SEE-- AND THIS IS KIND OF A PROMOTION IF ANY OF YOU ARE WORKING ON THESE TARGETS, STRONG INTERACTION OF APRIL WITH HIP 1ALPHA-- IT ONLY OCCURS IF YOU LOOK IN THE FAR RIGHT, IT OCCURS BOTH IN THE 1005, THE PRIMARY, AND ALSO ANESTHETIC, AND ONLY UNDER HYPOXIC CONDITIONS, AND ACTUALLY IN THESE CELLS, THIS IS MY POINT, IN THESE CELLS, IT DOESN'T INTERACT WITH NF KB. IT DOESN'T MEAN IT DOESN'T INTERACT WITH IT AT ALL, BUT WE CAN'T GET A PULLDOWN WITH IT. SO IF THERE IS AB INTERACTION, IT'S NOT AS MUCH AS IT IS-- YOU CAN TELL WITH HIP 1ALPHA, IT REALLY STRONG. SO THIS IS OUR-- THE WORLD ACCORDING TO APE, THE SIGNALING NODE, IF YOU WILL, AND YOU CAN START NOW TO LOOK AT COMBINATIONS BY LOOKING AT INTERACTIONS, WE KNOW APE INTERACTING WITH STAT3, WE HAVE A STAT3 INHIBITOR, DO YOU HAVE A HIF-1ALPHA INHIBITOR. SO I'M GOING TO SHOW YOU JUST A COUPLE EXAMPLES OF HOW IT CAN BE A HYPOTHESIS-DRIVEN COMBINATION APPROACH THAT WE THINK FOR TREATMENT OF CANCERS. SO THE FIRST APPROACH IS STAT3. MELISSA SHOWED STAT3 IS UNDER REDOX CONTROL BY APE REF-1 AND SHE'S BEEN WORKING WITH SOME PEOPLE IN CANADA FOR SOME STAT3 INHIBITORS BUT THEY'RE NOT FAR ENOUGH ALONG YET TO USE CLINICALLY. THERE IS A DRUG THAT IS AVAILABLE CLINICALLY FOR-- IT'S UPSTREAM OF STAT3, AND WE'VE PARTNERED THAT WITH 3330-- THIS IS DATA MELISSA HAD-- SORRY I'M GOING TO SKIP THROUGH THIS BECAUSE I'M RUNNING OUT OF TIME, JUST SHOWING THE IMPORTANCE OF APE INTERACTING WITH STAT3, WHICH LED US TO THE NEXT STEP WHERE WE WANTED TO COMBINE. AGAIN, WE REALLY WANT TO HAVE A STAT3 INHIBITOR, THERE ARE NONE PROVED YET SO WE WENT UPSTREAM OF THAT. IF YOU LOOK IN THE MIDDLE PANEL, THE MIDDLE GRAPH, WE GET A TRUE SYNERGISTIC EFFECT OF ADDING RUX PLUS 3330, BECAUSE YOU'RE REALLY GETTING IT AT TWO POINTS, YOU'RE BLOCKING THE PHOSPHORYLATION SIGNALING OF STAT3 AND YOU'RE BLOCKING THE REDOX ACTIVATION OF STAT3. YOU CAN THEN TAKE THIS MODEL AND START TO LOOK AT OTHER CANCERS AND OTHER PATHWAYS. HIF-1ALPHA, WE KNOW REF1 REGULATES IT, 3330 BLOCKS THAT. CA9IS-- INVOLVED REGULATING PH AND TUMOR CELLS FOR GROWTH. THERE'S A CLINICAL DRUG BEING TESTED IN CANADA, WHICH ISN'T SUPER EFFECTIVE IN ITSELF SO FAR, BUT WE APOLOGIZED BLOCKING AT TWO POINTS, ONE, BLOCKING THE PRODUCTION OF IT, AND TWO, BLOCKING WHATEVER'S MADE THAT YOU WOULD GET THIS COMBO EFFECT, SO WE INITIALLY DID THE KNOCK DOWN AND IT'S ONLY ON IN HYPOXIA, SO HYPOXIA-DRIVEN HIF1. KNOCKING DOWN OR GIVING OUR DRUG, WE GET A HUGE REDUCTION OF CF9THAT'S MADE, AND WHEN WE DO, OUR DRUG AND OUR DRUG PLUS SLC, YOU CAN SEE THERE'S A COMBO EFFECT. WE GET A LITTLE BIT OF CAF KILLING, IT LOOKS LIKE, BUT WE STILL IF YOU LOOK AT THE BOTTOM OF THE SLIDE, THE SECOND PANEL IN FROM THE LEFT, YOU CAN SEE YOU'VE REALLY WIPED OUT AND BY THE THIRD PANEL COMPLETELY WIPED OUT THE TUMOR CELLS. SO THIS AGAIN IS OUR HYPOTHESIS OF BLOCKING THE PATHWAYS-- THIS IS JUST THE-- OF THAT. I'M NOT GOING TO GO THROUGH THIS BECAUSE I MENTIONED THIS EARLIER BUT THIS HAS BEEN THE SAME SORT OF THING WHERE YOU CAN NOW PARTNER IT WITH-- 33. SO THIS ALLOWS US TO LOOK AT WHAT CANCERS WE NEED TO FOCUS ON. YOU START TO PARTNER UP NOW THE DIFFERENT MODELS AND THEN PARTICULARLY IN PATIENT TRIALS WITH THE RIGHT DRUG AND THE RIGHT COMBINATION AT THE RIGHT TIME. SO IN ABOUT THREE MINUTES, I'M GOING TO RUN THROUGH THIS AND THOSE OF YOU WHO ARE LOCAL HERE, IF YOU WANT TO TALK MORE ABOUT IT, I'D LOVE TO, BUT I HAVE TO GET TO THIS BECAUSE THIS IS ACTUALLY MORE REPAIR AND IT'S ACTUALLY MORE WHAT WE'VE BEEN FOCUSING ON LATELY. CHEMOTHERAPY-INDUCED PERIPHERAL NEUROPATHY. YOU TALK TO MOST PHYSICIANS AND THEIR PATIENTS HAVE THIS PROBLEM A LOT OF THE TIME AND THERE ARE NO APPROVED DRUGS FOR THIS. THE DRUGS-- PATIENTS COME OFF THEIR TREATMENT DUE TO PERIPHERAL NEUROPATHY EFFECTS, SOME OF IT'S SHORT TERM, SOME NEVER GOES AWAY. I'VE DONE LONG TERM STUDIES OF PATIENTS TREATED WITH CYST PLATINUM AND THEY STILL HAVE-- DECADES OUT FROM BEING TREATED AND THEY HAVE NEUROPATHIES. -- ALSO YOU SEE THIS, AND REMEMBER-- IF YOU REMEMBER THAT DATA BY THAT OTHER LAB SHOWING 3330 ENHANCED 5FU, KEEP THAT IN MIND AS I GO THROUGH THIS. THESE ARE SOME OF THE THINGS THAT PEOPLE TELL YOU WHAT IT FEELS LIKE WALKING ON HOT COALS, FINGER NAMES ON A CHALKBOARD. AND THAT'S CONSTANT. SO THESE ARE REALLY SYMPTOM MANAGEMENT BUT ALSO QUALITY OF LIFE BUT IT ALSO AFFECTS YOUR TREATMENT BECAUSE YOU COME OFF TREATMENT. SO WE'RE TOTALLY CONVINCED, BIASED, I GUESS YOU COULD SAY, THAT DNA DAMAGE IS REALLY A MAJOR COMPONENT OF PERIPHERAL NEUROPATHY, AND THIS IS THE SPINAL CORD SHOWN WITH PERIPHERAL NEURONS COMING OUT THAT GO TO YOUR HAND AND YOUR FEET, AND THE DORSAL ROOT BEGAN GLEE ON NEURON ARE OUT THERE SO THEY'RE LESS PROTECTED. WHAT YOU SEE IS A LOT OF DNA DAMAGE HAS BEEN SHOWN BY THE VARIOUS AGENTS, ROS PRODUCTION, MITOCHONDRIAL DNA DAMAGE, CHANGE IN PERMEABILITY, ET CETERA. SO I THINK THIS IS A RIPE AREA REALLY FOR THE DNA REPAIR FIELD AND PERIPHERAL NEUROPATHY. HUGE, HUGE UNMET NEED. WE'RE BIASED BECAUSE-- AND I THINK THIS IS SOMEWHAT STILL TRUE THAT VER IS ONE OF THE MAJOR PLAYERS IN THE NEURONS ALONG WITH NER. WE'RE STARTING TO SEE SOME CROSSTALK BETWEEN THESE PATHWAYS. WE'VE BEEN FOCUSING ON THE PLATINUMS BECAUSE THERE'S THREE REALLY NICE ONES YOU CAN WORK WITH. ALL THREE CAUSE NEUROPATHIES, CARBOPLATIN NOT AS MUCH. -- CAUSE SIGNIFICANT A OXIDATIVE DAMAGE IN ROS. CISPLATIN-- I ENCOURAGE THIS AS A NICE ASSORTMENT OF DRUGS WHERE YOU HAVE DIFFERENT STRAND CROSS LINKS IN DIFFERENT CONFIGURATION, SO YOU COMPARE THEM AGAINST EACH OTHER. WHAT WE'VE SHOWN ACROSS THIS PANEL HERE, I'M SORRY I'M GOING TO HAVE TO GO THROUGH QUICKLY IS, BASICALLY CGRP RELEASE IS RELEASED BY THE DORSAL ROOT BEGAN GLEANS. YOU CAN DO THIS IN VIVO AND YOU CAN ALSO DO THIS WITH CELLS IN CULTURE, AND THAT'S AN INDICATION OF THE DORSAL ROOT GLEE ON FUNCTION AND BASICALLY THE BOTTOM LINE IS CIS, OX AND CARBO-- I DON'T HAVE TIME TO GO THROUGH IT, BUT THAT'S DUE, WE THINK, TO THE OXIDATIVE DNA DAMAGE. IN THE LOWER RIGHT-HAND SIDE OF PANEL C, WHAT WE'RE SHOWING HERE IS IF YOU DO A KNOCKDOWN OF THE RAT APE BY RAT SIS AND YOU BRING IN THE HUMAN WHICH IS NOT AFFECTED BY THE RAT SI, YOU CAN NOW GET A NICE KNOCK DOWN OF APE WITH AN ADD-BACK, AND WHEN WE BROUGHT IN THE REDOX MUTANT, IT CORRECTED THE DEFECT IN THE-- RELEASE, BUT WHEN WE BROUGHT IN THE REPAIR MUTANT, IT DID NOT CORRECT IT. SO IT'S REALLY THE REPAIR FUNCTION OF APE THAT'S IN THE NEURONS. ONE OF THE THINGS WE SAW, WHEN YOU KNOCK DOWN APE, YOU HAVE A LONGER RETENTION OF THE PLATINUM EDUCT. WE KNOW IT DOES NOT WORK ON THAT, PER SE, AND SO WHAT IS GOING ON, AND WE LOOKED AT ADDING BACK WILD TYPE APE AND WE CAN GET THIS CORRECTION, IF YOU LOOK AT THE TOP PANEL, OVER TIME. WHAT WE THINK IS HAPPENING AND WHAT WE'RE GOING TO LOOK AT IN COLLABORATION WITH JOHN TURCHI IS WE THINK OXIDATIVE DNA DAMAGE IS AFFECTING THE RATE OF NER REPAIR. SO WE THINK NOW THERE'S A NICE CROSSTALK BETWEEN THE BER PATHWAY AND THE NER PATHWAY. ONE LAST THING I WILL SHOW YOU IS THAT WHEN WE DID IN VIVO EXPERIMENTS WITH OUR DRUG, 3330, AND WE LOOK FOR BLOOD FLOW WHICH IS AN INDICATION OF NEURONAL FUNCTION, WE CAN CORRECT IT IN THE RIGHT-HAND SIDE, WE CAN CORRECT THE DEFECT THAT YOU SEE WHEN YOU TREAT WITH CISPLATIN BY ADDING OUR DRUG. WHAT WE THINK IS HAPPENING IS IF YOU LOOK AT THE SQUARE RED BOX, IS THAT WE REALLY PREVENT DNA DAMAGE. NOW THIS IS JUST-- I KNOW SOME OF US LIKE THAT AND SOME OF US HATE THIS, WE'RE NOT SAYING WHAT KIND OF DAMAGE, BUT IT'S A MARKER FOR DAMAGE THAT GETS COMPLETELY PREVENTED WHEN OUR DRUG IS PRESENT WHEN WE'RE DOING THE TREATMENT, AT THE SAME TIME WE SEE A CHANGE IN PREVENTION AGAINST THE FUNCTIONAL END POINT OF THE IPN. SO WE'VE DONE THIS-- THIS IS WITH CISPLATIN-- WE JUST RECENTLY DID OX-- SO WE'RE VERY EXCITED THE DRUG IS NOT ONLY GOING TO HAVE A TUMOR EFFECT BUT IT'S GOING TO PROTECT THE NEURONS FROM BEING DAMAGED. HOW DOES THAT HAPPEN? WE DON'T KNOW EXACTLY. OUR WORKING HYPOTHESIS AND WE CAN TALK ABOUT IT, IS THAT THE DRUG IN THE NEURON-- THE NEURONS DO NOT KEEP DOWN AND LET THE-- SIT THERE-- YOU GIVE THEM ANY DAMAGING AGENT OR CISPLATIN CELLS AND YOU KNOCK APE DOWN, WE HAVE DYSFUNCTION AND SOME WILL DAY. WE THINK WHAT HAPPENS IS THE D. IT'S GOING TO APRIL, IT'S CAUSING A LOCAL INSULTING, BUT APE NOW IS FREED UP TO DO MORE REPAIR ACTIVITY. WE'RE TRYING TO PROVE THAT. WHEN WE TAKE-- FROM TREATED CELLS, BUT WE HAVE TO NOW FIGURE OUT THE EXACT MEASUREMENT. BUT THAT'S HOW WE'RE EXPLAINING IT RIGHT NOW, BUT I ADMIT WE'RE IN OUR INFANCY OF EXPLAINING IT, BUT THIS IS THE PHENOTYPE YOU GET. SORRY. SO VERY QUICKLY, IF I CONVINCE YOU OF NOTHING ELSE, I REALLY THINK WE HAVE TO, FOR APE AND ALL THE OTHER GENES, I THINK REPAIR JEANS, MULTIPLE ROLES AND THEY'RE NOT THE SAME. IT MAY HAVE SIMILAR ROLES BUT WHAT THEY END UP DOING FUNCTIONALLY IS DIFFERENT. I'M TRYING TO FIGURE-- FOR TRANSLATIONAL, I THINK IT'S BEEN VALIDATED FOR CANCER, THERE'S INDICATIONS FOR OTHER DISEASES THAT WE'RE WORKING ON, AND WE'RE PICKING THEM BASED ON THOSE PATHWAYS THAT ARE INVOLVED-- TRON DESCRIPTION FACTORS THAT APE REGULATES AND WE'RE GOING TO DO THE TRIAL, AND I THINK THIS NEURONAL PROTECTION, I THINK CAN BE PRETTY BIG. I HAVE ACTUALLY A SMALL GROUP, MY LAB AND THEN THE TOP RIGHT, WE DO A LOT OF COLLABORATION, WE LOVE TO COLLABORATE, SO IF ANY OF YOU ARE INTERESTED, WE HAVE A LOT OF THE DRUGS, IF ANYONE IS INTERESTED IN USING THE DRUG OR ANYTHING ELSE, WE'D LOVE TO DO THAT, BECAUSE THERE'S NOTHING MORE EXCITING THAN COLLABORATING WITH FOLKS LIKE YOURSELVES. SO SORRY I WENT LONG, BUT-- I CAN TAKE QUESTIONS NOW. [APPLAUSE] >> THANKS, MARK, FOR A REALLY INTERESTING TALK. FROM [INAUDIBLE] WE'RE GOING TO GO AROUND IF ANYONE HAS A QUESTION, ONE QUESTION PER SITE, AND EVERYBODY ELSE MUTE. >> CAN YOU HEAR ME? >> YES. >> GREAT TALK. I REALLY ENJOYED IT. JUST A VERY QUICK QUESTION. MOST OF THE DATA YOU PRESENTED WERE PATIENT-DERIVED XENOGRAPHS. DO YOU KNOW IF THIS WORKS IN A SYNGENEIC MODEL IN A MOUSE AND DO YOU KNOW IF THERE'S AN EFFECT ON THE IMMUNE SYSTEM IN TERMS OF PD1BLOCKADE OR ANYTHING LIKE THAT? >> THE IMMUNOTHERAPY QUESTION, YES. SO? >> THE IMMUNOTHERAPY QUESTION, YES. SO WE ARE JUST STARTING THE-- MODEL, YOU THERE ARE GENETIC MODELS FOR PANCREATIC CANCER. WE DO CELLS FROM THE GENETIC MOUSE MODEL, WE SEE THE SAME EFFECT. BUT THAT'S A GREAT QUESTION BECAUSE WE DO HAVE TO LOOK AT THE IMMUNE-- THERE'S DATA I HAVE THAT'S INVOLVED IN THE IMMUNE SYSTEM, SO WE'RE GOING TO BE DOING MORE OF THOSE. WE HAVEN'T DONE ANYTHING WITH PD1INHIBITORS OR ANYTHING LIKE THAT, BUT IF YOU'RE DOING IT OR SOMEONE IS DOING IT, WE'D LOVE TO DO THAT. >> OKAY, THANK YOU. >> [INAUDIBLE] >> HI, SCOTT WILLIAMS, NIEHS. I'M JUST WONDERING IF YOU COULD CLARIFY, SO THE REDOX CYCLING REACTION IN APE 1WITH SYS65, SO GOING FROM THE REDUCED TO THE OXIDIZED STATE, IT'S FORMING DISULFIDE BONDS, AND HOW DOES THE DRUG IMPACT THAT EXACTLY? IT'S A LITTLE HAZY ON THIS UNFOLDING REACTION. I JUST WANTED A LITTLE BIT OF ELABORATION ON THAT POINT, IF YOU CAN. >> SO THE DRUG 3330BINDS WITH-- 65AND BINDS BETWEEN 93AND 65OR 99AND 65, STARTS TO UNFOLD AND OPEN UP AND WE THINK EXPOSES IT THEN TO THE LOCAL CELLULAR ENVIRONMENT, WHICH CAUSE IT IS TO BECOME OXIDIZED. IT ALSO BLOCKS SOME OF THE THYROID OXIN RECHARGING, OTHERS HAVE SHOWN IT INVOLVED IN CONVERTING APE FROM THE OXIDIZED TO REDUCED STATE. >> I GUESS A FOLLOW-UP THEN, DO YOU THINK THAT THE REDOX ACTIVITY IS INVOLVED IN REGULATING ANY DNA REPAIR FACTORS? >> THAT'S A GOOD QUESTION. WE HAVEN'T LOOKED DIRECTLY, ANY OF THE DNA FACTORS THAT ARE OXIDIZED THAT NEED TO BE REDUCED? >> RIGHT. AFTER CC16789 UNDER GHOST-- TO INFLUENCE-- BETA INTERACTIONS? >> DON'T KNOW, BUT YOU WANT TO TEST IT, THAT WOULD BE GREAT. YOU KNOW, YOU COULD ALSO ENVISION THAT THE TRANSCRIPTION FACTORS ARE REGULATING THE REPAIR ENZYMES EXPRESSION, BUT WHEN WE DID THE KNOCK DOWN, WE NEVER SEE A LOT OF REPAIR PROTEINS CHANGE A LOT, AT LEAST AT THE RNA LEVEL. BUT THAT'S A GOOD EXPERIMENT. THAT'S INTERESTING. >> THANK YOU. GREAT TALK. >> THANKS. >> [INAUDIBLE] >> HAVEN'T DONE THAT, NO. WE HAVEN'T LOOKED AT TRANSLOCATION MUCH. DID SOME TRANSLOCATION EFFECT BUT HE USES LEVELS OF 33 PRETTY HIGH. LIKE I SAID, WE HAVE-- WE'RE HAPPY TO DO THAT. >> LET'S GO TO-- IN NCI. >> WE HAVE A QUESTION HERE. >> HI. THIS IS-- AT NCI. VERY NICE TALK, QUITE EXCITING DATA. AS YOU KNOW, ONE OF THE IMPORTANT END POINT IN PANCREATIC CANCER TREATMENT IS THE SURVIVAL, AND AS YOU MENTIONED EARLIER IN RESPONSE TO THE FIRST QUESTIONS THAT YOU ARE USING KPC MICE, SO DO YOU SEE ANY DIFFERENCE IN SURVIVAL USING THAT GENETIC ENGINEERED MOUSE MODEL? MY SECOND QUESTION IS THAT UP WITH OF THE MAJOR ISSUES IS THE DELIVERY OF DRUG TO THE TUMOR BECAUSE OF THE-- STROMA, SO HOW WILL YOU TACKLE THOSE PROBLEMS WITH ACCESSIBILITY OF DRUGS TO TUMORS? GREAT QUESTION. WE HAVEN'T DONE THAT EXPERIMENT YET. IT'S NOT THE MOST FAVORABLE, NOT THE EASIEST MODEL TO WORK WITH, YET ENOUGH MICE TO DO THE STUDY, SO WE'RE IN THE MIDDLE OF DOING THAT. WE'VE DONE THE KPC CELLS WHERE WE TOOK FROM KPC MICE AND PUT THEM IN A NORMAL MOUSE AND SAW AN EFFECT, BUT THAT'S NOT EXACTLY THE SAME, ALTHOUGH THE MICE DID HAVE A FULL IMMUNE SYSTEM. THE SECOND PART OF THE QUESTION ABOUT DELIVERY, AND LEAST IN OUR STUDIES, WE SEE THE DRUG GET TO THE TUMOR AND WE'VE DONE THAT IN OUR TUMOR AND OUR CAF MODEL, OF COURSE THAT'S NOT LIKE A FULL PANCREATIC TUMOR, SO ONE OF THE THINGS WE'RE WILLING TO DO, AS YOU KNOW WITH. PANC PATIENTS, WE HAVE PANC PATIENTS IN THE PHASE 1TRIAL, THEY WILL HAVE GONE THROUGH A LOT OF OTHER TREATMENTS ALREADY, AND THERE PROBABLY WON'T BE ANY TUMOR TO MEASURE IT IN. IF WE HAVE OTHER SOLID TUMORS AND WE CAN GET A NEEDLE BIOPSY OR SOMETHING LATER ON, WE COULD LOOK FOR A DRUG IN THE TUMOR. UNLESS YOU WERE DESIGNING IT SPECIFICALLY EARLY ON TO LOOK FOR THE DRUG IN THE TUMOR, IT WILL BE A LITTLE DIFFERENT IN PANC. GOOD QUESTION, THOUGH. THANKS. >> [INAUDIBLE] ANYONE ELSE? >> YES, MARK, THIS IS ANOTHER MECHANISM-RELATED QUESTION. HOW IS THE EFFECTIVENESS OF 3330 INFLUENCED BY THE PROTEIN'S INTERACTION WITH OTHER PROTEINS? FOR EXAMPLE, SO NOT THE TRANSLOCATION BUT HOW EFFECTIVE IS THE DRUG WHEN YOU HAVE THE INTERACTION VERSUS WHEN YOU DON'T HAVE IT? >> I'M NOT SURE I UNDERSTAND. >> YOU CAN ENGINEER-- THE DRUG HAVE ANY ADDITIONAL EFFECT UNDER THOSE CONDITIONS? >> WE HAVEN'T DONE THAT, NO. SO NO. >> BECAUSE IT SEEMS LIKE MPM1 WOULD INCLUDE INTERACTION WITH THE DRUG, RIGHT? IT'S BINDING ALONG EVIDENTLY THE SAME SURFACE NEARBY. >> WITHOUT DOING THE EXPERIMENT, I DON'T KNOW. IT'S A GOOD QUESTION. WE DON'T KNOW WHAT MPM1DOES IN THE CELLS WE'RE LOOKING AT, THE TUMOR TYPES, YOU KNOW, GOOD QUESTION BUT WE'D HAVE TO DO ANOTHER SERIES OF EXPERIMENTS. >> DIFFERENT ANGLE WOULD BE TO DO SOME-- WHERE IT TAKES THE PROTEIN TO THE CYTOPLASM. >> YEP, WE COULD DO THAT TOO. I WILL TELL YOU-- >> OR-- CAN DO IT ANYWAY. >> I THINK THAT'S THE WAY TO DO IT, YEAH. NO, THAT'S A GOOD POINT. THAT'S A REALLY GOOD EXPERIMENT TO DO. GOOD QUESTION, BRUCE. WE HAVEN'T DONE THAT. SOMETHING TO LOOK AT. WE DO HAVE SOME EVIDENCE NOW THAT IN AN INFLAMMATION INDUCED AML MODEL, NOT NECESSARILY THE TRANSLOCATION AND MPM-RELATED, BUT THAT WITH THE CERTAIN EPIGENETIC PHENOTYPE THAT THESE ARE CELLS TO GO ON TO DEVELOP AML IN A MOUSE MODEL, THAT IF WE GIVE OUR DRUG, WE PREVENT THOSE CELLS FROM CONVERTING INTO FULL BLOWN LEUKEMIA, AND THAT'S A PAPER THAT WILL BE SUBMITTED SOON. NOT EXACTLY ANSWERING YOUR QUESTION BUT VIAL TO LOOK IN THOSE AMC CELLS TO SEE-- IT WOULD BE SOMETHING CARLO WOULD LOOK AT, THE SAME MPM SORT OF INTERACTION. >> THANKS. >> THANKS, BRUCE. >> NOW WE GO TO KENTUCKY, ARE YOU THERE? >> WE'RE THERE. THIS IS DAVE. GREAT TALK. >> GO AHEAD, DAVE. BRUCE, CAN YOU MUTE, AND DAVE-- >> OKAY, THANK YOU.